IMMUNOENZYMATIC ASSAY THAT MEASURES THE EXPRESSION OF MURINE HISTOCOMPATIBILITY ANTIGENS IN MACROPHAGES AND LYMPHOCYTES

A convenient-simple, sensitive, rapid and reproducible enzyme immunoas say to measure H-2 particulated and solubilized cellular antigens is d escribed. Cellular antigens were measured by ELISA through the binding of specific biotinylated antibodies and streptoavidin-peroxidase conj ugate to cells in suspension. Endogenous peroxidase activity of activa ted cells was inhibited by addition of sodium azide and H2O2 in acid c onditions. The assay proved capable of distinguishing between two cell lines (EL-4/H-2 b and P815/H-2d) and even between the cells of three congenic mouse strains (BALB/B, H-2b , BALB/c, H-2d, and BALB/K, H-2k) and was sensitive to as few as 2.5 x 10(4) cells/well. Results were c omparable to those obtained with FACS. An inhibition version of this a ssay was found to be very useful for the detection of H-2 antigens pre sent in whole antigen cells extracts. (C) 1993 Wiley-Liss. Inc.